Nanomedicine: from high tech to global health
Transcript of Nanomedicine: from high tech to global health
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Nanomedicine: from high tech to global health
Robert K. Prud’homme, Brian Johnson,Walid Saad, Ying Liu, Marian Gindy, Stephanie Budijono, Margarita Herrera-Alonso,Varun Kumar,Suzanne D’Addio, Robby Pagels,
Chet Markwalter, Brian WilsonDept. Chemical Engineering
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Support: NSF, NIH, Gates Foundation, BASF, Evonik, Merck, J&J, GSK, Celator, Optimeos; Nanomedicine for BBB-crossing in CNS oncologic pathologies MAECI PROJECT 2019-2021
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Next Generation Nanoparticles (NPs)
Tumor vasculature
Nanoparticles
11 µm
“…over the next few years some of the complex theranostic strategies published rampantly in chemistry journals will fall out of contention…For me, something that’s too difficult to make or too complex to sustain in large-scale production is not what we are interested in.”
J.Janijic C&EN Sept 26,2011.
“…over the next few years some of the complex theranostic strategies published rampantly in chemistry journals will fall out of contention…For me, something that’s too difficult to make or too complex to sustain in large-scale production is not what we are interested in.”
J.Janijic C&EN Sept 26,2011.
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Block copolymer directed rapid precipitation
Nanoparticle formation by Flash NanoPrecipitation
Non solvent
Copolymer Solute Solvent
DeadMicelle
PROTECTEDNANO
PARTICLE
tmix
Micellizationtagg
tngNucleationand Growth
Johnson, Prud’homme AICHE J (2003), Liu, Prud’homme, Fox, Chem Engr. Sci. (2008)
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• Control super -saturation by changing solvent quality or solute concentration.
• Higher super saturation leads to smaller particles
Control of particle size
0 100 200 300 400 500 60050
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300 1:1 H2O:THF 2:1 H2O:THF 3:1 H2O:THF 5:1 H2O:THF
Parti
cle D
iam
eter
(nm
)
Average Re
1. Mixing intensity
2. Super-saturation
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NP Formation: PS to PLA NPsWant to shift from model polystyrene (PS) to biocompatible polylactic acid (PLA). How does size depend on species in core?
• What is in the core doesn’t matter
• The ratio of core to BCP matters, Ccore /CBCP
• The total mass matters
PS-b-PEG
PS-b-PEG
PLA-b-PEG
PLA-b-PEG
Pagels Nano Letters 18 1139-1144 (2018)
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NP Formation: PS to PLA NPsWant to shift from model polystyrene (PS) to biocompatible polylactic acid (PLA). How does size depend on species in core?
• What is in the core doesn’t matter
• The ratio of core to BCP matters, Ccore /CBCP
• The total mass mattersPagels Nano Letters 18 1139-1144 (2018)
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Drugs ConsideredClofazimine
Ref: Design and Solidification of Fast-Releasing Clofazimine Nanoparticles for Treatment of Cryptosporidiosis”. Molecular Pharmaceutics. 14(10) 3480-3488 (2017); “Rapid Recovery of Clofazimine-loaded Nanoparticles with Long-term Storage Stability as Anti-Cryptosporidium Therapy”. ACS Applied Nano Materials. 1(5) 2184-2194 (2018)
OZ439
Ref: “Encapsulation of OZ439 into Nanoparticles for Supersaturated Drug Release in Oral Malaria Therapy”. ACS infectious diseases. 4(6) 970-979 (2018); “Spray drying OZ439 nanoparticles to form stable, water-dispersible powders for oral malaria therapy”. Journal of Translational Medicine (2019), just accepted
Lumefantrine
Ref: “Amorphous Nanoparticles by Self-assembly: Processing for Controlled Release of Hydrophobic Molecules”. Soft Matter. 15 2400-2410 (2019)
Delamanid
Ref:https://app.box.com/s/lvghy956h7pmzvdd1nlj9zzeotabijke
Ecumicin
Ref:https://app.box.com/s/5lggnm4tv84kcb0cry0tscby9oszzqzi
TBA 354
Ref:https://app.box.com/s/57cromgxhg8uyqiz4zegtafwqbclg3nk
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Clofazimine – Zein/casein
Direct spray dryingredisperses to NP
Mol Pharm (2017)
40X higher dissolution in GF
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Long-term Zein Clofazimine NPs
Day FaSSIF FeSSIF
0 5.4 ug/mL 5.4 ug/mL
1 4.5 ug/mL 4.8 ug/mL
3 4.7 ug/mL 5.9 ug/mL
7 6.2 ug/mL 5.6 ug/mL
14 4.6 ug/mL 5.1 ug/mL
• Open vial: 75 % RH, 50 C
• Have gone to 28 da +
• FeSSIF: influence was notsignificant
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0 50 100 150 200 250 300 350 400
Rele
ase
(%)
Time (min)
FaSSIF Day 0
Day 1
Day 3
Day 7
Day 14
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0 50 100 150 200 250 300 350 400
Rele
ase
(%)
Time (min)
FeSSIF
Fasted Intestinal Fluid
Feasted Intestinal Fluid
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siRNA for gene silencing
•siRNA complex cleaves mRNA and stops protein expression
•Problem is delivery
+
buffersiRNA
ethanolcationic lipid
structural lipid
+
+++
++ +
+
+
•Successful prep of 100 nm PEG protected siRNA
•Stable and transfect efficiently
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1 mL batch
1.5 L/min 5 L/min
CIJConfined Impinging Jet
MIVMMulti Inlet Vortex Mixer
Micro-MIVMµ−Multi Inlet Vortex Mixer
Markwalter, et al JOVE 143 (2019), J Pharm Sci 107 (2018); Feng, J Trans Med (2019)
Gates: 3700 tons/yr
Four Scales of CIJ & MIVM
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Equivalent NP formation vs Reynolds No.
Similar NPs produce from batch (1 ml scale) to 5 L/min
Feng, et al. J. Translational Medicine (2020)
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Active targeting
Small molecule:(mannose, folate, LHRH)
• Conjugate targeting group to block copolymer• Do hard chemistry and characterization on
conjugate• Quantitatively assemble targeted NP with
control on % targeting groups• Conjugate chemistry: click chemistry, maleimide,
carbodiimide, carbamate
Gindy, Biomaterials (2008)Akbulut Adv Fn Mat (2009)
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Variable domain antibody targeting
ELN
–N
anop
artic
les 2
013
-000
48
Blank Cells
W26A-NPs (300 ug/mL)
•14K fibrinogen peptide scaffold (Centryin)
•Pre functionalize BCP•Assemble NP
Control (PEG NP)
5% ligand targeted
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Inverse Flash NanoPrecipitation (iFNP)
Precipitation process for the encapsulation of hydrophilic therapeutics
iFNP uses a non-polar antisolvent to produce NPs with a water-
soluble core:
iFNP has been demonstrated on a variety of water-soluble molecules:
All ≥ 50% loading Other encapsulated molecules include: polysaccharides, insulin,
ovalbumin, peroxidase, Trp cage , mRNA
Pagels & Prud’homme, J. Cont. Release (2015)Markwalter, et al. AAPS (2020)
20
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Unique Technology Capability: RNA Encapsulation
mRNA has been encapsulated (50% loading) into inverted nanoparticles by iFNP
Cationic lipids are not required for encapsulation of RNA –different from currently available technologies
Model RNA used: RNA from Torula utilis, Mr 5,000-8,000 Da (15-25 nucleotides)
0
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Inte
nsity
Diameter, nm
50 wt% RNA
PK1 avg: 40 nmPDI: 0.24
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0.5
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230 250 270 290 310 330 350
Abso
rban
ce, A
.U.
Wavelength, nm
Solvent blankRNA controlNPs
95% encapsulation efficiency
RNA control – no
polymer
RNA encapsulated
by iFNP
RNA precipitate (left) and nanoparticles (right) in
CHCl3
Particle size distribution Encapsulated RNA is protected from extraction into an
aqueous phase
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Unique Technology Capability Co-Encapsulation
RNA (33 wt% loading) and a model protein (33 wt% loading)
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Inte
nsity
Diameter, nm
RNA + HRPNPs
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225275325375425Wavelength, nm
RNA +HRPRNA
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225275325375425
Abso
raba
nce
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Brine phase
Organic NP phase
Nanoparticle Size Distribution Nanoparticle Encapsulation Efficiency
RNA
HRPHorseradish peroxidase,
44 kDa
PLA-b-PAsp
Unencapsulated Encapsulated
Nanoparticle Composition, wt%
Encapsulation Efficiency
RNA HRP Polymer RNA HRPRNA NPs. 50% -- 50% 92% --HRP NPs. -- 50% 50% -- 99%
RNA + HRP NPs.33.3% 33.3% 33.3% 93% 99%
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Cu-64 PET/CT imaging allows tracking of biodistribution in vivo
23
Neutral PEG coating leads to tumor accumulation
3 hours post injection 24 hours post injection
Negative coating promotes immune cell interactions
24 hours post injection
For Discussion with Spark Therapeutics Only Please Do Not Distribute
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Summary ([email protected])
Flash NanoPrecipitation (FNP) Polymer-directed, rapid precipitation process Hydrophobic drugs, and drugs made hydrophobic by ion pairing Scaleable: mg to 1000 kg Enables targeting incorporation and imaging (PET, fluorescence, MRI)
Inverse Flash NanoPrecipitation (iFNP) Polymer-directed, two step rapid precipitation process Hydrophilic peptides, proteins, mRNA, DNA High loading (20-50%), high encapsulation efficiency (80-99%) Makes nanoparticles, or microparticles for depot delivery (2 mo release)